6-methyl-9alpha-fluoro derivatives of hydrocortisone and cortisone and 21-esters thereof



W Nam 6-METHYL-9oz FLUORO DERIVATIVES F HYDRO- CORTISONE AND CORTISONE AND ZI-ESTERS .-THEREOF George B. Spero, Kalamazoo, Mich., assign'or to The Upjohn Company, Kalamazoo, Mich., a corporation of Michigan No Drawing. Application March *6, 1957 Serial No. 644,214

Claims. (Cl. 260397.45)

The present invention relates to steroids and is more particularly concerned with 6-methyl-9a-fiuorohydrocortisone, 6-!1'16thYI-9a-fiUOI'OCO1tiSO116, the 21-esters thereof and a process for the production thereof and the production of the A -analogue, 1-dehydro-6-methyl-9u-fluorohydrocortisone.

The present invention is a continuation-in-part of application Serial No. 624,965, filed November 29, 1956, and now abandoned.

The compounds and the process of the present invention are illustratively represented by the following sequence of formulae:

2,928,51 Patented Mar. 15, 1960 wherein X is a halogen of atomic number between seventeen and 37, inclusive, wherein Ac is the acyl radical of an organic carboxylic acid, preferably a hydrocarbon carboxylic acid containing from one to twelve carbon atoms, inclusive, and wherein R is selected from the group consisting of hydrogen and acyl, the acyl radical being defined as above.

The process of the present invention comprises: dehydrating a 6-methylhydrocortisone 21-acylate (I) illustratively with an acid, such as hydrochloric acid, sulfuric acid or with thionyl chloride or preferably, with an N- haloamide such as N-bromoacetamide and followed by treatment with anhydrous sulfur dioxide, to obtain the corresponding 6 methyl -17a,2l-dihydroxy-4,9(11)-pregnadiene-3,20-dione 2l-ester (II); treating the thus-obtained 6-methyl 17a,21 dihydroxy-4,9(11)-pregnadiene-3,20- dione 2l-acy1ate or the free alcohol, i.e., 6-methyl-17a,21- dihydroxy-4,9(11)-pregnadiene-3,20-dione which can be produced by hydrolysis of the ester under conventional conditions, with a hypohalous acid such as hypochlorous or hypobromous acid, preferably formed in situ, results in 6-methyl-9a-halo 11fi,17oc,21 trihydroxy-4-pregnene- 3,20-dione 21-acylate or the free alcohol (III). Treatment of III with a mild base suchas anhydrous potassium acetate yields the epoxy compound, 6-methyl-9,8,1118- oxido-17a,21-dihydroxy-4-pregnene-3,20-dione 21-acylate or, respectively, the unesterified product, represented by Formula IV. Treatment of the epoxy compound (IV) with hydrogen fluoride or with hydrogen fluoride release ing agents provides the physiologically active fluoro de-v rivative of Formula V, i.e., 6-IIIGthYI-9oc-fillOIO-l1fl,17oc, 21-trihydroxy-4-pregnene-3,20-dione 21-acylate or the free alcohol, 6-methyl-9ot-fluoro-hydrocortisone. Oxidation of the esterified compound (V) with chromic acid provides the 6-methyl-9u-fluoro-17a,2l-dihydroxy-4-pregnene-3,1l,20-trione 21-acylate (VI). Hydrolysis of the ester VI with a base provides the free alcohol, G-methyl- 17 12,2 l-dihydroxy-4-pregnene-3, 1 1,20-trione 6-methyl-9ocfluorocortisone). Subjecting 6-methyl-9inc-fiuorohydrocortisone or the ester thereof to dehydrogenation either by micro-organisms such as Septomyxa or with selenium dioxide produces the A analogue, 1-dehydro-6-methyl-9w: fiuorohydrocortisone, or esters thereof. The same compounds V and VI may also be obtained by the alternate process shown in Example 13.

It is an object of the instant invention to provide the new adrenocortical hormones, 6-methyl-9a-fluorohydrocortisone, 6methyl-9u-fluorocortisone and the 21-acylates thereof, in particular in the 6a-epimeric form. It is another object of the instant invention to provide a method for the production of 6-methy1-9oc-fluorohydrocortisone, 6-methyl-9afluorocortisone, the ZI-esters thereof and 1-dehydro-6-methyl-9u-fiuorohydrocortisone.

vide the intermediates for the production of these adreno- I comically-active compounds, such as 6-methy1-17a,21-di-' hydroxy-4,9 1 i -pregnadiene-3,20-dione 2l-acylate, 6- methyl 9 l l )-oxido-170;,2l-dihydroxy-4-pregnene-3,20- dione 2l-acylate, the 9a-chloroand 9ot-bromo-6methyl l15,l7 2l-trihydroxy-4-pregnene-3,ZO-dione 2i-acylates and the free alcohols thereor". Other objects of this invention will be apparent to those skilled in the art to which this invention pertains.

The novel 6-methyl-9es-fluorohydrocortisone, the 6- methyl-9a-luorocortisone and the Zl-esters thereof, in particular the 6wepimers thereof, i.e., 6a-methyl-9txfluorohydrocortisone and 6a'methyl-9a-iluorocortisone, are very active adrenal cortical hormones. The glucocorticoid activity of the novel compound, 6ot-methyl- Qa-fluorohydrocortisone surpasses the glucocorticoid activity of the natural hormones, hydrocortisone and cortisone, and the novel compound possesses in addition antiinflammatory activity of an extremely high order. The 6-methyl-9a-fluorohydrocortisone, the 6-methyl-9ot-fluorocortisone and the 2l-esters thereof are thus useful in parenteral and'topical compositions and may be given as tablets for oral use in combination with such binding materials and carriers as polyethylene glycol 4000 or 6000, lactose, sucrose, and the like. Especially useful for this purpose is the 6o:-methyl-9a-iluorohydrocortisone and esters thereof. In topical application the substances can be used as ointments, lotions, jellies, crearns, suppositories, bougies, aqueous suspensions, and the like. Instead of the 6a-methyl-9a-fluorohydrocortisone or 60:- methyl-9m-fluorocortisone, the 6p-epimers thereof can be used in therapeutically equivalent amounts to give the r same final results.

The 6ot-methyl-9a-tluorohydrocortisone is also an important intermediate in the production of the extremely active 1-dehydro-6a-methyl-9a-fiuorohydrocortisone as shown in Example 15. The l-dehydro-6a-methyl-9afluorohydrocortisone was found to have a subcutaneous glucocorti coid activity between 108 and 127 times that of hydrocortisone, while the oral glucocorticoid activity was found to be 191 times that of hydrocortisone.

The starting material of the instant invention are the 6ot-methylhydrocortisone ester as described in Preparations 1 through 10. Instead of the 6ct-methylhydrocortisone, the 6B-epimer can be used to give the corresponding 6/3-methyl-9ot-fiuorohydrocortisone and the 6fimethyl-9ot-fluorocortisone.

In carrying out the process of the instant invention,

6-inethyl-1-15,l7ot,2l-trihydroxyi-pregnene 3,20 dione- ZI-acylate (6-methylhydrocortisone 2l-acylate) is dehydrated to the corresponding 21-acylate of 6-methyl- 17h,21-dihydroxy-4,9(1l) -pregnadiene 3,20 dione by methods known in the art, for example, by a dehydrating agent such as phosphorus oxychloride, thionyl chloride, hydrochloric acid or sulfuric acid and acetic acid or by pyrolysis as shown by US. Patents 2,640,838 and 2,640,839. In the preferred embodiment of the present invention the dehydration is eifected by reacting the llfl-hydroxy compound with a carboxylic acid N-haloamide or N-haloimide in an organic base and treating the thus produced intermediate 1l-hypohalite with dry sulfur dioxide in an organic base. As reagents for the production of an ll-hypohalite, the acid N-haloamide or acid N-haloimide are used wherein the halogen has an atomic number from 17 to 53, inclusive, preferably chlorine or bromine. Examples of such compounds are N-chloroacetamide, N-brornoacetamide, N-chlorosuccinimide, N-bromosuccinimide, N-iodosuccinimide, 3- bromo-5,5-dimethylhydantoln and ll,3-dibromo-5,5-dimethylhydantoin, and the like. Ordinarily an amount in excess of a molar equivalent, calculated on the starting llfi-hydroxy steroid, is employed. The organic base employed as solvent in the production of the ll-hypohalite are tertiary amines wherein the amino nitrogen is a member of an aromatic ring, for example, the pyridines, that is, pyridine, alkyl-pyridines, piccoline, lutidine, collidine, con'yrine, parvuline, or the like, bi

' thirty degrees centigrade).

lower fatty amides such as formamide, methylformamide and dimethylformamide. The base is preferably employed in a large molar excess, calculated on the starting llfi-hydroxy steroid, for example, ten molar equivalents per mole of steroid, and is preferably the sole reaction solvent. The reaction to produce an ll-hypohalite is generally conducted under anhydrous conditions, preferably, containing less than 0.1 molar equivalentof water calculated per mole of steroid. Large proportions of water decrease the yield. The temperature of the reaction is between minus forty and plus seventy degrees centigrade, the lower limit being determined by the solubility of the reaction and solvents and the upper limit being determined by the amount of side reaction which normally accompanies the reaction involving any halo compounds at higher temperatures. Ordinarily, room temperature (twenty to thirty degrees centigrade) is preferred for-convenience and because of the consistently high yields of the desired product which are obtained at this temperature. A reaction period between five minutes to three hours is usually employed; at higher temperaturesabove thirty degrees centigradeshort reaction times are sutncient to efiect completion of the reaction.

The thus produced 6-methyl-l1,8,l7a,2l-trihydroxy4- pregnene-3,20-dione llfi-hypohalite, ZI-acylate is then treated with anhydrous sulfur dioxide in the presence of an organic base as described hereinbefore. The anhydrous sulfur dioxide can be in the form of gaseous. or liquid sulfur dioxide or in the form of a material which in situ produces sulfur dioxide, for example, alkali metal hyposulfite. The reaction temperature ranges substantially within minus forty and plus seventy degrees centigrade and preferably room temperature (twenty to The thus obtained product a 6-methyll7ot,2l -dihydroxy-4,9 l 1 -pregnadiene-3,20- dione 21-acylate is isolated from the reaction mixture by conventional means such as extraction after the reaction mixture has been poured into excess of water. Organic water-immiscible solvents such as ether, chloroform, methylene chloride, carbon tetrachloride, ethyl acetate, benzene, hexanes, or the like, are used for the extraction. The thus obtained extracts are conveniently washed, dried and thereupon evaporated to give the crude 6- methyl-17a;1-dihydroxy-4,9( 11)-pregnadiene-3,20-dione 2l-ester which is purified by conventional means such as recrystallization or chromatography, as deemed necessary.

The thus obtained 6-methyl-17ot,21-dihydroxy-4,9(ll)- pregnadiene- LZO-dione 21-acylate is converted too-methyl 90:3- halo 1l;8,l7a,21 trihydroxy 4-pregnene 3,20-dione ZI-acylate by adding a hypohalous acid such as hypochlorous or hypobromous acid. The hypohalous acid is usually added by reacting an N-haloacidamidecr an N-haloacidimide with an acid to produce the hypohalous acid in situ. In the preferred embodiment of the invention, the steroid, a 6-methyl-l7a,2l-dihydroxy- 4,9(11)-pregnadiene'3,20-dione 21-ester is dissolved in an organic solvent such as methylene chloride, tertiary butyl alcohol, dioxane, tertiary amyl alcohol, or the like, and reacted at room temperature with the hypobromous or hypochlorous acid releasing agent in the presence of an acid. Such hypohalous acid releasing agents include the N-bromoacetamide, the N-chloroacetamide, the N-bromosuccinimide, the N-iodosuccinimide, the N-chlorosuccinimide, or the like, in the presence of water and an acid such as perchlo-ric acid, dilute sulfuric acid, or the like.

; The reaction is usually carried out at room temperature,

between fifteen to thirty degrees centigrade, however, lower or higher temperatures are operative for the process. The hypohalous acid. releasing agent is generally used in one molar or slightly increased, for example, 25 percent increased, amounts compared to mole of steroid. A large excess of the hypohalous acid releasing agent while operative is undesirable, since the excess of hypohalou'sacid has a tendency to react on otherpositions of the molecule. The reaction period is rather short and may vary between about four to five minutes to one hour. At the end of the reaction time excess of hypohalous acid is destroyed by the addition of sodium sulfite or other sulfites or hydrosulfites. The thus produced product, a 6 methyl 9oz halo 11B,l7a,21 trihydroxy 4- pregnene-3,20-dione 21-acylate, wherein the halogen atom is of atomic weight between 33 and 130 (atomic number 17 to 53), is isolated from the reaction mixture by adding excess of water and extractingthe compound with organic solvents or by recovering the precipitated compound through fitration. The crude product thus obtained may be recrystallized from organic solvents, such as acetone, Skellysolve B hexane hydrocarbons or the like to give pure 6-methyl-9a-halol 1B,17cz,2 l-trihydroxy-4-pregnene- 3,20-dione 21-acylate.

The ester thus obtained can be hydrolyzed in an acidic or neutral medium to give the free triol, 6-methyl-9a-halo- 11B,17c ,21-t1ihydroxy-4-pregnene-3,ZO-dione, which can be reesterified by known methods of esterilication of 21- hydroxy steroids of the pregnane series.

' Oxidizing the 2l-ester of a 6-methyl-9u-halo-l1,3,l7a,21-

trihydroxy-4-pregnene-3,20-dione 2l-acylate with chromic acid produces the corresponding pharmaceutically active 6 methyl 9a halo 170:,21 dihydroxy 4 pregnene 3,11,2-0-trione ZI-acylate which by hydrolysis in an acidic or neutral medium gives the free triol 6-methyl-9a-halo- 17oz,2 1-dihydroxy-4-pregnene-3 ,l 1,20-trione.

In order to obtain the 9tz-fiuoro compounds, the 9 8,11 [3- epoxy intermediates of the before-mentioned compounds, 6 methyl 9 6,115 oxido 17a,21 dihydroxy 4 pregnene-3,20-dione 2l-esters, are prepared. In carrying out this reaction a 6-methyl-9a-halo-l1p,17a,2l-trihydroxy-4-pregnene-3,ZO-dione 21-ester wherein the halogen atom is of atomic number seventeen to 35, inclusive, is heated in solution with a mild base, and preferably in the absence of water to avoid hydrolysis of the ester groups.

The bases useful for the cyclization include anhydrous potassium acetate, sodium bicarbonate, sodium acetate, or the like, with potassium acetate preferred. Solvents such as methanol, ethanol, acetone, tertiary butyl alcohol, or the like, may be used. The reaction time is between one half hour and 24 hours; generally a period between three and twelve hours is sufficient. The thus obtained 6- methyl 96,115 oxido l7a,21 dihydroxy 4 pregnene-3,20-dione 21-acylate is isolated from the reaction mixture by diluting the reaction mixture with excess water and filtering the product when crystalline, or

by extracting with methylene chloride or other Waterimmiscible solvents such as ether, Skellysolve B hexanes, pentanes, benzene, ethyl acetate, methylene chloride, chloroform, carbon tetrachloride, or the like. Evaporation of the solvent of the extracts produces the 6-methyl- 95,115 oxido 17a,2l dihydroxy 4 pregnene -3,20 dione 2l-acylate.

The thus obtained 6-methyl-9;3,1lB-oxido-lhJl-dihydroxy-4-pregnene-3,20-dione 2l-acylate is thereupon reacted with hydrogen fluoride either as gas cr'liquified at low temperature or with 48 percent hydrofluoric acid in solution. As solvents for this reaction methylene chloride, ethylene dichloride, tetrahydrofuran, chloroform, carbon tetrachloride, or the like, is useful. The reaction may be carried out at room temperature (twenty to thirty degrees centigrade) or, in the preferred embodiment of the invention, in tetrahydrofuran at low temperature, e.g., zero to minus eighty degrees centigrade with stirring. The period of reaction is from one to 24 hours with a period from one to twelve hours usually suflicient. After the reaction is terminated, the mixture is poured into water and neutralized with a dilute base, e.g., sodium or potas sium' hydroxide or carbonate or a bicarbonate such as sodium bicarbonate, potassium bicarbonate, or the like. The reaction mixture is then extracted with a waterimmiscible solvent such as methylene chloride-theorganic layer is separated from the water mixture, washed with water, dried and evaporated to give the crude 6-methylc fiuoro 11(3,l7a,2l trihydroxy 4-pregnene 3,20 dione 2l-acylate. The thus obtained crude compound may be purified through recrystallization or chromatography.

The 6-methyl-9a-fluoro-1 1 3, l7u,21-trihydroxy-4-pregnene-3,20-dione 21-esters obtained by this process can be hydrolyzed with a base or acid in conventional manner to give the 6-methyl-9a-fluoro-l1B,l7a,2l-trihydroxy-4- pregnene-3,20-dione which can be re-esterified, if desired, with acyl halides or acid anhydrides, in pyridine solution at room temperature to give other 2l-esters thereof.

Oxidation of the 6-methyl-9 x-fluoro-11,6,17a,2l-trihydroxy-4-pregnene-3,20-dione 2l-acylate with chromic acid produces the. corresponding 6-methyl-9a-fiuoro-17a,21- dihydroxy-4-pregnene-3,11,20-trione 2l-acylate which can be hydrolyzed illustratively with a base, e.g., with sodium carbonate in ethanol in a nitrogen atmosphere to give the free diol, 6-methyl-9a-fiuoro-17a,2l-dihydroxy-4-pregnene-3 ,l1,2-0-trione.

The dehydrogenation of 6-methyl-9a-fluoro-1l,8,17a,21- trihydroxy-4-pregnene-3,20-dione to obtain the A -analogue thereof is carried out either by fermentative or chemical dehydrogenation. Microorganisms such as Septomyxa, Corynebacterium, Fusarium, and the like, are used under fermentation conditions well known in the art (e.g., US. 2,602,769) and furthermore illustrated by Example 14 herein. In case of the use of Septomyxa it was found to be advantageous to use with the substrate and medium a steroid promoter, such as progesterone, 3-ketobisnor-4-cholen-22-al, B-ketobisnorcholenic acid, 115,21-dihydroxy-1,4,l7(20)-pregnatrien-3-one, and the like. The chemical dehydrogenation can be carried out with selenium dioxide as illustrated in detail by Example 15.

The following preparations and examples are illustrative of the process and products of the present invention, but are not to be construed as limiting.

PREPARATION 1 5a,6oz-0xid01 1,8,1 7a,21-trihydroxyalZ0pregnane-3,20- dione 3,20-bis-(ethylene ketal) To a solution of 0.901 gram of 11B,l7a,2l-trihydroxy- 5-pregnene-3,20-dione 3,20-bis-(ethylene ketal) in eighteen milliliters of chloroform was added a solution of 331 milligrams of perbenzoic acid in 5.19 milliliters of chloroform. The resulting solution was allowed to stand in the refrigerator (ca. four degrees centigrade) for a period of 24 hours and thereupon at room temperature for an. additional period of 72 hours. The reaction solution was then washed with five percent sodium bicarbonate solution and water, dried over anhydrous sodium sulfate and evaporated to dryness to give 1.031 grams of crude solid. Recrystallization from acetone gave 431 milligrams of material of melting point 230 to 247 degrees centigrade. The mother liquor, after evaporation to dryness, was dissolved in methylene chloride and chromatographed over 25 grams of acid washed alumina. The, column was developed with three fractions each of methylene chloride plus five, ten, fifteen, twenty, 25 and fifty percent acetone, acetone, and acetone plus five percent methanol. The acetone plus five percent methanol eluate gave an additional 279 milligrams of the high melting product. The high melting material, 5a,6 xoxido-l 1,6,17a,2l-trihydroxyallopregnane-3,ZO-dione 3,20 bis-(ethylene ketal) was three times recrystallized from acetone and methanol to give a pure product of melting point 263 to 268 degrees centigrade. Other eluate fractions of lower melting point contained the 518,6[3-isome'r thereof. M

In the same manner as shown in Preparation 1, other- 5a,6a oxido 1l;8,17a,21 trihydroxyallopregnane-3,20 dione 3,20-bis-(alkylene ketals) can be prepared by re acting hydrocortisone diketals, wherein the ketal group asses has been formed, by reacting the steroid 3,2G-dione with ethylene, propylene, 1,2-, 1,3-, or 2,3-butylene glycol or pentane,'hexane, heptane, or octane-diols wherein the alcohol groups are in vicinal positions such as 1,2, 2,3, 3,4, or the like, or separated by one carbon atom such as 1,3,, 2,4, 3,5, and the like, with an organic peracid such as performic, peracetic, perbenzoic, monoperphthalic acid, or the like. For the purpose of this invention, starting compounds having the ethylene ketal groups are preferred, since these ketals are generally more easily prepared in high yield than ketals produced by the reaction of the 3,20-diketo compounds with higher alkanediols.

PREPARATION 2 5 a,] 1,3,1 70;,21 -tetrahydroxy-6fi-methylall0pregnane- 3,20-dione 3,20-bis-(ethylene keml) A solution of 1.115 grams of Soc,6a-oxido-1l,6,17,21- trihydroxyallopregnane 3,20 dione 3,20-bis-(ethylene ketal) in 165 milliliters of tetrahydrot'uran (the tetrahydrofuran being dried through distillation over lithium aluminum hydride) was added dropwise to a solution of 95 milliliters of methyl magnesium bromide in ether (the magnesium bromide having a four molar concen tration). To this mixture was added 575 milliliters of benzene and the reaction mixture was thereupon allowed to stir and reflux for 26 hours. After cooling, the reaction mixture was poured into 700 milliliters of iced, saturated ammonium chloride solution, stirred for a period of thirty minutes, and the benzene layer separated from the aqueous layer. The aqueous phase was extracted with three 20ll-miililiter portions of ethyl acetate and the extracts added to the benzene layer. The combined benzene-ethyl acetate solution was thereupon washed with water, dried over anhydrous sodium sulfate and evaporated to dryness to give 1.314 grams of crude solid. Trituration of this material with ether left 1.064 grams of crystalline product of melting point 221 to 230 degrees. Recrystallization of this material gave a,1i 8,l7rx, 2l-tetrahydroxy-fl-methylallopregnane-3,20-dione 3,20- bis-(ethylene ketal) of melting point 228 to 233 degrees and rotation [a1 minus eleven degrees in chloroform.

Analysis.-Calcd. for (3 11 0 C, 64.70; H, 8.77. Found: C, 64.29; H, 8.69.

PREPARATION 3 50:,1 1b,] 70:,21-tetrahydroxy-6fi-ethylallopregnanc- 3,20-dione 3,20-bis-(ethylene keml) In the same manner as shown in Preparation 2, 511,60:- oxido-l 15,17u,21-trihydroxyallopregnane3,ZO-dione 3,20- bis-(ethylene hotel) was reacted with ethyl magnesium bromide in ether solution to give the corresponding 50:, 1l 3,l7oz,Zl tetrahydroxy 6B ethylallopregnane 3,20- dione 3,20-bis-(ethylene ketal).

In the same manner as shown in Preparations 2 and 3, other 512,1 1f3,1704,21-tetrahydroxy-6fi-alkylallopregnane-3, -dione 3,20-bis-(ethylene ketals) are prepared by reacting the corresponding 5a,6e.-oxido-11;8,l7e,21-trihydroxy-allopregnane-3,ZO-dione 3,20-bis-(ethylene lcetal) with a metal alkyl or metal aryl more specifically an alkyl metal halide such as a Grignard reagent, for example, methyl, ethyl, propyl, isopropyl, butyl, iso-butyl, pentyl, hexyl, and phenyl magnesium bromides and iodides or cadmium alkyl and calcium alkyl and phenyl bromides or iodides. Representative 6fi-alkylated allopregnanes thus prepared include:

5u,1lfi,l7oz,2l tetrahydroxy 6/3 propylallopregnane- 3 ,20-dione 3,20-bis- (ethylene ketal) 5,0t,1lB,l7oz,21 tetrahydroxy 65 butylallopregnane- 3,20-dione 3,20-bis- (ethylene ketal),

5a,l1fl,17oa,2l tetrahydroxy 61S isobutylallopregnane- 3,20-dione 3,20-bis-(ethylene ketal),

5a, 1 1fl,l7a,2l tetrahydroxy 6,3 pentylallopregnane 3,20-dione 3,20-bis- (ethylene ketal),

511,1 1p,17u,21 A tetrahydroxy 6n hexylallopregnanc i 3,20-dione 3,20-bis-(ethylene ketal) 5u,1lfi,l7u,21 tetrahydroxy 6B phenylallopregnane 3,20-dione 3,20-bis-(ethylene ketal), and the like.

PREPARATION 4 A solution was prepared containing 468 milligrams of 5ot,l1fl,l7a,21'-I6ll3hYd1OXY 6,6 methyiallopregnane- 3,20-dione 3,20-bis-(ethylene ketal), 38 milliliters of methanol and 7.7 milliliters of 2N'sulfuric acid. This solution was refluxed for a period of thirty minutes, then neutralized with five percent dilute sodium bicarbonate solution (about milliliters) and concentrated under reduced pressure at 55 degrees centigrade to about 35.

milliliters of volume. A product crystallized upon cooling and was recovered by filtration. This product was recrystallized from acetoneSkellysolve B hexanes to give an analytical pure sample of 50:,11B,l7a,21-tetrahydroXy 6B-methylallopregnane-3,ZO-dione of meltingpoint 240 to 244 (decomposition) and rotation [al plus forty degrees in dioxane.

Analysis.-Calcd. for C22H3406Z C, H, 8.69. Found: (3,6684; H, 8.86.

PREPARATION 5 5 a,1 15,1 7a,21-tetrahydroxy-6B-etlzylallopregnane 3,20-di0ne In the same manner as shown in Preparation 4,

5a,1l 8,17u,21 tetrahydroxy 6p ethylallopregnane- 3,20-dione 3,2G-bis-(ethylene ketal) was hydrolyzed with 5a,11B,17a,21 tetrahydroxy 6,8 propylallopregnane- 3,20-dione,

5oz,l1j3,17ot,21 tetrahydroxy 6p butylallopregnane- 3,20-dione,

5m,11,3,17a,21 tetrahydroxy 6e isobutylallopregnanc- 3,20-dione,

5a,llfl,l7oz,21 tetrahydroxy 6;? pentylallopregnane 3,20-dione,

5oz,llfi,17x,2l tetrahydroxy 6B hexylalloprcgnane- 3,20-dione,

5a,llfi,l7ot,2l tetrahydroxy 6,8 phenylallopregnane- 3,20-dione, and the like.

PREPARATION 6 6ot-methylhydroc0rtis0ne milligrams was recrystallized from acetone-Skellysolve B hexanes to give in two crops 249 milligrams of 6cr-methylhydrocortisone melting between 184 and 194 degrees centigrade. An analytical sample was prepared melting at 203 to 208 degrees centigrade and consisting of 6a: methylhydrocortisone.

g. Analysis-Calcd. for C H O C, 70.18; H, 8.57. Found: C, 70.32; H, 8.50.

The mother liquors contained besides 6a-methylhydrocortisone, substantial amounts of 6/3-methylhydrocortisome which can be isolated by recrystallization, papergram, countercurrent procedures and other means known in the art.

.Esterification of 6ot-methylhydrocortisone with acetic anhydride in pyridine at room temperature yielded 604- methylhydrocortisone 21-acetate of melting point 213 to 214 degrees centigrade.

Analysis.Calcd. for Found: C, 68.60; H, 8.41.

PREPARATION 7 6/8-methylhydr0c0rtis0ne 45x 0 c, 68.87; H, 8.19.

A solution was prepared containing 27.5 grams of a,l1,8,17a,21 tetrahydroxy 6,8 methylallopregnane- 3,20-dione in 6500 milliliters of ethanol denatured with methanol. The solution was freed of air oxygen by bubbling oxygen-free nitrogen through it for a period of fifteen minutes. To this solution was added a similarly air oxygen-free prepared solution of one-tenth normal sodium hydroxide (235 milliliters). The solution was allowed to stand at room temperature (about 22 to 24 degrees centigrade) in an inert nitrogen atmosphere for a period of twenty hours and was then acidified with fourteen milliliters of acetic acid. The thus obtained acid solution was evaporated at about fifty to sixty degrees centigrade in vacuo, the thus produced residue dissolved in 200 milliliters of ethyl acetate and 200 milliliters of water, the water layer separated from the organic layer and discarded, the organic layer washed with 350 milliliters of five percent aqueous sodium bicarbonate solution, then three times with water and thereupon dried over anhydrous sodium sulfate and concentrated to a volume of 180 milliliters. After cooling the 180 milliliters of solution in a refrigerator (about five degrees centigrade), the solution was filtered giving 11.9 grams of material. This material was redissolved in 500 milliliters of ethyl acetate, the ethyl acetate solution was concentrated to 150 milliliters, refrigerated as before to give 6.15 grams of crude 6fl-methylhydrocortisone of melting point 220 to 223 degrees centigrade.

Recrystallization of the crude 6B-methylhydrocortisone three more times from ethyl acetate gave an analytical sample of 6B-methylhydrocortisone with melting point 223 to 227 degrees centigrade, rotation [otJ plus 105 degrees in acetone; ultraviolet absorption A1 3 ethanol 243 m a 14,500

Analysis.--Calcd. for C H O Found: C, 70.54; H, 8.91.

PREPARATION 8 6oi-ethyllzydr0cortis0ne C,70.l7; H, 8.57.

Age? 243 614, 525

In the same manner dehydratin with an alkali metal hydroxide in alcoholic solution other 50c,l1B,17a,2'-.

tetrahydroxy-6B-alkylallopregnane-3,20-diones produced the corresponding 6a-alkyland 6et-aryl11,8,17ot,21-trihydroxy-4-pregnene-3,20-diones such as 6a propylhydrocortisone, 6ot-butylhydrocortisone, 6a-isobutylhydrocortisone, 6a-pentylhydrocortisone, -6ot-hexylhydrocortisone, 6a-phenylhydrocortisone, and the like.

1 0 PREPARATION 9 Goa-methylhydrocbrtisone acetate A mixture was prepared containing one gram of 6amethylhydrocortisone in five milliliters of pyridine and five milliliters of acetic anhydride. The mixture was maintained at room temperature (22 to 24 degrees centigrade) for a period of six hours, thereupon poured intomilliliters of ice water and the resulting aqueous mixture extracted with three 25-milliliter portions of methylene chloride. The combined methylene chloridesolutions were washed, dried over sodium sulfate and evaporated and the thus obtained residue recrystallized three times from acetone-Skellysolve B hexanes to give pure 6a-methylhydrocortisone 21-acetate (6ot-methyl-11 3, 17cc dihydroxy 21-acetoxy-4-pregnene-3,20-dione) of melting point 213 to 214 degrees centigrade.

PREPARATION 10 yielded 6u-methylhydrocortisone 21-benzoate.

(g) 6a-methylhydrocortisone with phenylacetyl chloride yielded a-methylhydrocortisone 21-phenylacetate.

(h) u-ethylhydrocortisone with acetic anhydride yielded 6ot-ethylhydrocortisone acetate.

In a manner similar to Preparations 9 and 10, other starting materials can be made by esterifying 6-alkylhydrocortisone or 6-arylhydrocortisone in pyridine solution with acyl halides or acid anhydrides. Starting materials thus prepared include the acetates, propionates, butyrates, isobutyrates, valerates, isovalerates, hexanoates, heptanoates, octanoates, benzoates, phenylacetates, B-cyclopentylpropionates, phenylpropionates, laurates, hemisuccinates, tartrates, maleates, toluenesulfonates, and the,

like of -alkylhydrocortisone and fi-arylhydrocortisone wherein the alkyl group is methyl, ethyl, propyl, butyl, pentyl, or hexyl and the aryl group may be phenyl or the like. v

EXAMPLE 1 6 ot-methyl-I 7e,21-dihydr0xy-4,9 (11 -pregnadiene-3,20-

dione 21 -acetate To a solution of 8.5 grams of 6a-methyl-1lfi,17u,21- trihydroxy-4-pregnene-3,ZO-dione 21-acetate (6a-rnethyl-" hydrocortisone 21-acetate) in 42.5 milliliters of pyridine was added 5.63 grams of N-bromoacetamide. After standing at room temperature for a period of fifteen minutes, the reaction solution was cooled to five to ten degrees. centrigrade and, with shaking, sulfur dioxide gas was passed over the surface until the solution gave no color with acidified starch-iodide paper. During the addition of sulfur dioxide gas, the reaction became warm. The temperature was kept under thirty degrees centigrade by external cooling and by varying the rate of sulfur dioxide addition. Thereafter to the reaction mixture 400 milliliters of ice water was added and the resulting precipitate collected by filtration. This material was recrystallized from acetone-Skellysolve B hexanes 'to give 5.78 grams of 6a-methyl-17a,21-dihydroxy-4,9( 1 l -pregnadi-. ene-3,20-dione 21-acetate of melting point to 169 de gr'ees centigrade. The mother liquors were evaporated to a partly crystalline residue which weighed 1.82 grams. This material was dissolved in methylene chloride and 11 chromatographed on 75 grams of Florisil. The column was eluted with. 2000 milliliters of 8% acetone-92% Skellysolve B and 1000 milliliters of 15% acetone-85% Skellysolve B. The eluant was collected in ZOO-milliliter fractions. Fractions 3 to 7 were combined, evaporated, and the residue recrystallized from acetone-Skellysolve B hexanes to yield 0.88 gram of 6a-methyZ-17 21-dihydroxy-4,9( 1 1 -pregnadiene-3,2 -dione 2 l-acetate of melting points 169-172 degrees centigrade. The combined yield therefore was 6.66 grams or 82.4 percent. An analytical sample obtained by Crystallization from acetone-Skellysolve B hexanes gave 6a-methyl-17u,21-dihydroxy4,9(l1)-pregnadiene-3,20dione 21-acetate of melting point 175-176 degrees centigrade and rotation [al plus 91 degrees in chloroform.

Analysis.Calcd. for C H O C, 71.94; H, 8.05. Found: C, 71.75; H, 7.71.

EXAMPLE 2 6a-methyl-9ix-brom0-113,17a,21-trihydr0xy 4 pregrtene- 3,20-diorze ZI-acetate (6a-methyl-9a-bromohydrocorlisone 21 -acetate) milliliters of water was added and the reaction mixturev was concentrated to a volume of about 500 milliliters under reduced pressure at about fifty degrees centigrade. At this point crystallization started. The concentrate was cooled in an ice bath and while stirring 500 milliliters of water was added. After stirring for a period of one hour, the crystalline product was isolated by filtration, the crystals were washed with water and airdried to give 6.88 grams (98.9% yield) of 6a-methyl-9abrorno-l lfi,17a,21-trihydroxy-4-pregncne-3,20-dione 21- acetate (6a-methyl-9ot-bromohydrocortisone 21-acetate) of melting point 159 to 161 degrees centigrade (with decomposition). An analytical sample was obtained from dilute acetone of melting point 153-155 and rotation [aJ plus 148 degrees in chloroform.

Analysis.Calcd. for C H O Br: Br, 16.07. Found: Br, 16.01.

EXAMPLE 3 6a-methyl-9BJ1fl-oxido-1 7a,21 -dihydroxy-4-pregnene- 3,20-di0ne 21 -acetate 1 To a solution of 6.78 grams of 6ei-methyl-9a-bromo 1 1B, l7a,2 l-trihydroxy-4-pregnene-3,20dione 21-acetate in .175 milliliters of acetone was added 6.78 grams of potassium acetate and the resulting suspension was heated under reflux for a period of seventeen hours. The mixture was then concentrated to approximately sixty milliliters of volume at reduced pressure on the stearn bath and thereupon diluted with water and extracted with methylene chloride. The methylene chloride extracts were combined, washed with water, dried overanhydrous After stirring'the reaction mixture for fifteen.

12 material was used without further purification in the next step. An analytical sample of 6a-methyl-9fi,11,6-oxido 17ot,2l-dihydroxy-4-pregnene-'3,20-dione 2l-acetate, obtained by recrystallization of a portion of the peak fraction from acetone-Skellysolve B hexaues, had a melting.

point of 180 to 182 degrees centigrade and rotation [mi of plus degrees in chloroform.

Analysis.-Calcd. for C24H32OqZ Found: C, 69.41;H, 7.93.

EXAMPLE 4 hydrofuran and then a solution of 500 milligrams (0.0012" of 6 x-methyl-9{5,1 1,8-oxido-1 7 x,2 l-dihydroxy-4- mole) preguene-3,20-dione Ill-acetate (combined chromatographic fractions in two milliliters of methylene chloride). The steroid solution was rinsed in with an additional one milliliter of methylene chloride. The light red colored solution was then kept at approximately minus thirty degrees centigrade for one hour and at minus ten degrees for two hours. At the end of this period it was mixed cautiously with an excess of cold sodium bicarbonate solution and the organic material extracted with the aid of additional methylene chloride. The combined extracts were washed with water, dried over anhydrous sodium sulfate and concentrated to approximately 35 milliliters of volume. This solution was ehromatographed over forty milliliters of Florisil anhydrous magnesium silicate. The column was diluted with 400 milliliters of seven percent acetone-93% Skeliysolve B hexanes, 500 milliliters of 10% acetone-90% Ske1lycentigrade and rotation [(1.11 plus 113 degrees in acetone.

Analysis.Calcd. for C H O F: C, 66.03; H, 7.62; F, 4.35. Found: C, 65.69; H, 7.49; F, 4.29.

EXAMPLE 4a 6ot-methyl-9ot-flu0r0-11,8,17u,21-trihydr0xy 4 pregnene- 3,20-dione 21-acetale (Alternate procedure) A solution was prepared of 250 milligrams of 6a-methy1-9fl,1lfi-oxido-17a,21-dihydroxy-4-pregnene-3,20 dione Zl-acetate in five milliliters of methylene chloride and thereto was added one milliliter of 4S percent'solution of aqueous hydrogen fluoride. The two-phase mixture was stirred for a period of twenty hours, then diluted with fifteen milliliters of methylene chloride and'carc- 7 fully poured into forty milliliters of water conatining 3.5

grams of sodium bicarbonate. After shaking to neutralize the excess hydrogen fluoride, the methylene chloride was separated and the water phase was extracted with more methylene chloride. The combined methylene chloride solution (about milliliters) was dried over anhydrous sodium sulfate, diluted with 25 milliliters of ether and chromatographed over twenty grams of Florisil synthetic magnesium silicate. The column was eluted as follows: I

13 TABLE 1 Fraction No. Solvent 1 (100 milliliters) 2-6 (40 milliliters each) 7-16 10 milliliters each) 17-21 (40 milliliters each) 22-26 (40 milliliters each) 27-30 (40 milliliters each) Skellysolve B hexane plus acetone (12%).

Skellysolve B hexane plus acetone (15%).

Skellysolvc IB hexane plus acetone (20 Skellysolve B hexane plus acetone (25 Skellysolve B hexane plus acetone (50%).

EXAMPLE 5 6a methyl 9oz fluoro 170:,21 --dihydroxy-4-pregnene- 3,11,20-trine 2] -acemte (6a-methyl-9a-flu0r0cortis0ne 21 -acezate) A solution was prepared containing in one milliliter of acetic acid fifty milligrams of 6ot-methyl-9a-fiuoro- 11,8,17a,21-trihydroxy-4-pregnene-3,20-dione 2 l-acetate, twenty milligrams of chromic anhydride and one drop (approximately fifty milligrams) of water. This mixture was shaken several times at room temperature and allowed to stand for four hours. Thereafter it waspoured into ten milliliters of water and refrigerated for twenty hours at about five degrees centigrade. The steroid which separated from the aqueous mixture was collected on filter paper and recrystallized three times from acetone to give 6a-methyl-9a-fluoro-17a,21-dihydroxy-4-pregnene- 3,11,20-trione 21-acetate (6a-methyl-9a-fluorocortisone 21-acetate) In the same manner as given in Examples 1 through but using as starting material the corresponding 6;?- isomers are obtained 6,6-methyl-9u-fluoro-1lfi,l7a,21-trihydroxy-4-pregnene-3,ZO-dione 21-acetate and 6fi-methyl- 90c-fl1101O-17ot,2I-dihYdlOXY-4-PI6gn6Hfi-3,11,20-t1i0n6 21-. acetate. Using instead of the acetates, other esters as starting materials in the series exemplified by Examples 1 through 5, such as propionate, butyrate, isobutyrate, valerate, benzoate, hexanoate, heptanoate, octanoate, phenylacetate, phenylpropionate, laurate, or the like of 6aor 6B-methyl-hydrocortisone yields the corresponding esters of 6ocor 6B-methyl-9a-iiuorohydrocortisone and -cortisone. Instead of the 6a-methylhydrocortisone, 6aalkyland 6-arylhydrocortisones can be used to give by the sequence of steps illustrated in Examples 1 through 5 the corresponding 6a-alkyl-9ot-fluorohydrocortisone esters and the ll-keto analogues, i.e., 6a-alkyl-9a-fluorocortisone esters and 6u-aryl-9a-fluorocortisone esters, wherein the alkyl groups can be ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, hexyl, and the aryl group can be phenyl, benzyl, or the like.

EXAMPLE 6 6 a-methyl-Qa-fluoro-l 1 3,] 7a,21-trihydroxy-4-pregriene- 3,20-dione (6a-methyl-9ot-fluorohydrocortisone) Three and one quarter (3.25) grams of 6a-methyl-9a-' Metliylene chloride-ether 14 I sixty-clegree-centigrade water-bath; Thereupon 250miili liters of water was added and the mixture chilled. The crystalline product was collected on a filter, washedwith water and dried to give 2.43 grams (83%) of crude 6mmethyl-9a-fluorohydrocortisone. Recrystallization of the crude material from methanol and Skellysolve B gave pure 6a-methyl-9ot-fluorohydrocortisone of melting point 228 to 230 degrees centigrade and rotation [al plus 112 degrees in acetone. p 4

Analysis.-Calcd. for C H O F: C, 66.98; H, 7.92; F, 4.82. Found: C, 67.20; H, 8.01; F, 5.47. w

An additional amount of product (0.24grarn, 8.2%) of 6-methyl-9a-fiuorohydrocortisone was obtained by. saturating the filtrate with sodium chloride, extracting withmethylene chloride, drying the extracts over sodium sulfate, evaporating and recrystallizing the thus obtained residue from benzene-methanol.

EXAMPLE 7 6a-methyl-9a-fluor0-1 711,21-dihydroxy-4-pregnahe- 3,11,20- tri0ne (6a-methyl-9ot-fluorocortisone) In the same manner given in Example 6, hydrolyzing with sodium carbonate in ethanol solution in a nitrogen atmosphere 6a 7 methyl 9oz fluoro 17a,21-dihydroxy-4- pregne'ne-3,11,20-trione 21-acetate yielded 6ot-methyl- 9a-fiuoro-17a,2l-dihydroxy-4-pregnene 3,20 dione (6amethyl-9a-fluorocortisone).

EXAMPLE 8 6oc-methyl-9a-flu0ra-11B,17a,21 trihydroxy 4-pregnene-' 3,20-dione 21-propi0nate (6mmethyl-9a-fluorohydrocortisone 21-pr0pi0nate) .I

EXAMPLE 9 6u-methyl-9u-flu0r0-11,8,17tx,21 trihydroxy 4-pregnev emixture was allowed to stand overnight for a period of.

eighteen hours and was thereupon diluted with fifty milliliters of water. The water solution was extracted with three ten-milliliter portions of methylene chloride, the.

methylene chloride fractions combined, washedlwith water, dried over anhydrous sodium sulfate andevapo rated to give a residue. This residue was recrystallized; from methanol to give 6u-methyl-9a-fluoro-11B,l7u,21-"

trihydroxy-4-pregnene-3,20-dione 21-benzoate.

EXAMPLE 10 6ot-methyl-9a-fluoro-11,B,17a,21-trihydr0xy 4 pregnene- 3,20-di0ne 21 -hemisuccinate (6u-methyl-9a-flu0r0hydrocortisone 21 -hemisuccinate) A solution was prepared containing 0.5 gram- 0f. succinic anhydride, 0.1 gram of 6a-methyl-9a-fiuoro- 11,8,17u,21-trihydroxy-4-pregnene-3,20-dione in five milliliters of pyridine. This solution was allowed tostand overnight for a period of twenty hours, was thereupondiluted with water and the mixture refrigerated and-- filte ed. The pr c p h s c ed on filt pepe was;

The solution was allowed to" recrystallized two times from methanol to give 6amethyl-9a-fiuoro 1l;8,l7a,21 trihydroxy 4 pregnene- 3,2'O-dione 21-hemisuccinate.

EXAMPLE 11 6u-mezhyl-9u-flu0r0-115,17u,21-trihydr0xy 4 pregnene- 3,20-dz'one 21-hemisuccinate sodium salt (ore-methyl- 9a-fluor0hydr0c0rtisone 21 -hemisuccinate sodium salt) Sodium hydroxide solution (0.1 Nomial) was slowly added to a stirred solution of 100 milligrams of 60:- methyl-9a-fluoro 11fi,17a,21 trihydroxy 4 pregnene 3,20-dione ZI-h'emisuccinate, dissolved in two milliliters of acetone, until the pH rose to about 7.4. During the addition of sodium hydroxide solution, five milliliters of water was also added. The solution was then concentrated at'25 degrees centigrade under vacuum to remove the acetone. The resulting aqueous solution of 6ocmethyl-9a-fluoro- 1lfi,17oc,21 trihydroxy 4 pregnene- 3,20-dione ZI-hemisuccina-te sodium salt was filtered, freeze-dried and recrystallized to give pure Six-methyl- 9u fiuoro-l1 8,17u,21-trihydroxy-4-pregnene 3,26 dione ZI-hemisuccinate sodium salt. I

EXAMPLE 12 In the same manner given in Examples 8 through 10,

reacting in pyridine solution at room temperature (twenty to thirty degrees centigrade):

(a) 6a-rnethyl-9a-fiuorohydrocortisone with butyric anhydride yielded 6a-methyl-9m-fluorohydrocortisone 21- butyrate.

-(b) 6u-methyl-9a-fluorohydrocortisone with valeric.

anhydride yielded 6oomethyl-9a-fluorohydrocortisone 21- valerate. 1

(d) 6a-methyl-9a-'fluorohydrocortisone with phenylacetyl chloride yielded 6a-methy1-9a-fluorohydrocortisone ZI-phenylacetate.

(e) 6a-methyl-9a-fluorohydrocortisone with phenylpropionyl bromide yielded 6u-methyl-9a-fluorohydro cortisone 2l-phenylpropionate.

(j) 6d-methyl-9a-fluorohydrocortisone with [i-cyclopentylpropionyl chloride yielded 6a-methyl-9a-luorohydrocortisone 21-(B-cyclopentylpropionate) (g) 6e-methyl-9a-fluorocortisone with propionic anhydride yielded 60t-IllthY1-9d-fillOIQCOI'tlSOHG 21-propionate.

(h) 6a-methyl-9wfiuorocortisone with valeric anhydride yielded 6tat-methyl-9a-fiuorocortisone 21-valerate.

(1') 6oz -methyl -91: -iluorocortiso'ne with phenylacetyl chloride yielded 6a-rnethyl-9a-fluorocortisone 21-phenylacetate.

(k) 60: -methyl 9 x -fluorocorti'sone with undecylenyl chloride yielded 6a-methyl-9a-fluorocortisone 2l-undecylenate. V

In a manner similar to Examples 8 through and 12, other esters of 6os-methyl9a-fluorohydrocortisone and 6 t-methyl-9a-iluorocortisone are prepared by reacting these steroids with the anhydride or acyl halides or organic carboxylic acids, particularly hydrocarbon carboxylic acids containing from one to twelve carbon atoms, inclusive. Representative 2l-esters thus prepared include in particular, besides the before-mentioned examples, the butyrates, isobutyrates, valelrates, isovaierates, hexanoates, heptanoates, octanoates, benzoates, phenyla'ce'tate's, phenylpropionates, fi-cyclopentylpropionatc's, tertiary butyiacetates, toluates, Z-furoate's, benzenesulfonates, toluenesulfonates, and the like of 6amethyl-9a iiuorohydrocortisone and 6umethyl-9a-fiuorocortisone. In the same manner 2l-esters of the 6,8-

methyl epimers and the '2l-esters of the 6m and ofi-alkyh and aryl analogues of these compounds can be prepared,

Treating the 6-methyl-9m-halohydrocortisones and -cortisones, wherein the halogen atom is chlorine,

bromine or iodine, with an acylating agent selected from halides or anhydrides of organic carboxylic acids, especially hydrocarbon carboxylic acids containing from one to twelve carbon atoms, inclusive, preferably at room temperature in pyridine solution produces the corresponding 21-esters of 6-methyl-9a-halohydrocortisone and 6-methyl-9a-halohydrocontisone and 6-methyl-9ahalocortisone.

The starting 6a-methyl-9a-halohydrocortisones and -cortisones wherein the halogen is chlorine, bromine or iodine are produced by procedures known in the art, such as reacting the 6u-methyl-17u,2l-dihydr0xy-4,9(11)- pregnadiene-S,20-dione 21-ester (of Example 1) with N- bromo, N-chloroor N-iodosuccinimide in the presence of an acid as shown in Example 2 to give the corresponding 6 methyl-9a-halo-11 3,170:,2l-trihydroxyl-pregnene- 3,20-dione Zi-ester. Mild acid hydrolysis with dilute sulfuric acid at room temperature produces the corresponding free triol, 6-methyl-9u-halohydrocortisone.

Oxidation of the 6-rnethyl-9a-halo-l1;3,17o:,21-trihydroxy-4-pregneue-3,ZO-dione 21-ester with chromic anhydride yields the 6methyl-9a-halocortisone 21-ester which by acid hydrolysis as mentioned above gives the free alcohol, 6methyl-9u-halocortisone.

EXAMPLE 13 6a-methyl-9a-flu0r0hydrocortisone from QOL-flUOIDhYdIOCOVIiSOI'ZE (a) 9a-FLUOROHYDROCORTISONE-3,20-BIS(ETI-IYLENE KETAL) To a solution of ten grams of 9a-fluorohydrocortisone acetate in 400 milliliters of ethylene glycol was added 300 milligrams of para-toluenesulfonic acid The solution was refluxed at a temperature of to degrees centigrade at two to three-milliliters pressure. The distillation was continued for a period of three to three and one half hours and during this period approximately milliliters of distillate was removed gradually. Thereafter the solution was cooled and SOOmilliliters of water was added. The mixture was then made alkaline with sodium bicarbonate and extracted three times with a total of 300 milliliters of methylene dichloride. The organic layer was washed with Water, dried over anhydrous sodium sulfate, a few drops of pyridine added, and the solution chromatographed on 800 grams of Florisil anhydrous magnesium silicate. The following fractions of Weight in Milligrams Fraction Solvent Fractions 13 to 15 were combined, evaporated and the residue recrystallized from ethyl acetate containing pyridine and dried to give 540 milligrams of 9ot-flll010- hydrocortisone-3,ZD-bis-(ethylene ketal) of melting point 187.5 to 189 degrees centigrade. It should be noted that in this ketalization operation the 21-ester group is saponified. Two recrystailizations from ethyl acetate gave pure 9a-fluorohydrocortisone 3,20-bis-(ethylene keta'l) of melting point 203 to 205 degrees Centigrade.

17 A nalysis e-Calcd. r c n o m C, 63.94; H, 8.16.

Found: C, 64.23; H, 8.06.

a) 50,6tz-OXIDO-9a. -FLUORO 11B,17a,21 rnmrnnox'r- ALLOPREGNAINE 3,20 DIONE 3,20-BIS-(E'I'HYLENE KETAL) To 508.5 milligrams of 9a-fluoro-1l)3,17a,21-trihydroxy-S-pregnene-3,20-dione 3,20-bis-(ethylene ketal),

dissolved in fifty milliliters of ethyl acetate, was added 1.3 millimoles of p'erbenzoic acid, dissolved in three milliliters of ethyl acetate. After two days of standing 1.06 mole equivalents of perbenzoic acid had been consumed according to iodometric titration. The reaction mixture was then washed twice with saturated sodium bicarbonate, once with water, dried over anhydrous sodium sulfate and chromatographed over fifty grams of Florisil anhydrous magnesium silicate, taking fractions of 100 milliliters each as follows in Table III:

Fractions 12 through 15 were combined and crystallized from ethyl acetate containing pyridine to give 210 milligrams of pure 5a,6a-oxido-9a-fiuoro-l1fl,l7u,21-trihydroxyallopregnane-3,20-dione 3,20-bis-(ethylene ketal) of melting point 203 to 205 degrees centigrade. v

Analysis.Calcd. for C H F: C, 64.08; H, 7.96. Found: C, 64.23; H, 8.06.

(0) a,11B,17a,21-TETRAHYDROXY 6a METHYL-Qa-FLU- OROALLOPREGNANE-3,20-DIONE 3,20-BIS-(ETHYLENE KETAL) A solution of 1.16 grams of 5a,6a-oxido-9a-fluoro- 11p,l7a,21 trihydroxyallopregnane 3,20 dione 3,20- bis-(ethylene ketal), dissolved in 165 milliliters of tetrahydrofuran (distilled previously over lithium aluminum hydride), was added dropwise to a four molar solution of 95 milliliters of methyl magnesium bromide in ether. Thereafter 575 milliliters of benzene was added and the reaction mixture was allowed to stir and reflux for a period of 24 hours. After cooling the reaction mixture was poured into 700 milliliters of ice and saturated ammonium chloride solution, stirred therein for a period of thirty minutes and the benzene layer separated. The aqueous layer was extracted with three ZOO-milliliter portions of ethyl acetate and the extracts added to the benzene. The combined benzene-ethyl acetate extracts were washed with water, dried over anhydrous sodium sulfate, and concentrated to give 1.36 grams of crude solid 5 11,1 1,8,17a,21-tetrahydroxy-6ot-methyl-9a-fluoroallopregnane-3,20-dione 3,20-bis-(ethylene ketal). This crude material was triturated with ether and thereupon recrystallized from ethyl acetate to give pure 501,115.17 ca, 21 tetrahydroxy 6oz methyl 9oz fiuoroallopregnane- 3,20-dione 3,20-bis-(ethylene ketal).

(d) 5a,11B,17a,2l-TETRAHYDROXY-Ga-METHYL-Qa- FLUOROALLOPREGNANE-3,20-DIONE ZO-dione 3,20-bis-(ethylene ketal) in 38 milliliters of methanol was added 7.7 milliliters of two normal sul- 18 furic acid. After refluxing the mixturejor a period of one hour the solution was made neutral .with, dilute sodium bicarbonate, concentrated under reduced pressure at 35 to 40 milliliters and refrigerated between zero to five degrees centigrade. After standing at this temperature for a period of eighteen hours the mixture was filtered and the precipitate thus obtained recrystallized from ethyl acetate Skellysolve B-hexanesv to give pure 5a,11B,17u,2l tetrahydroxy 6a methyl 9a fluoroallopregnane-3,2O-dione.

(6) Ga-METHYL-Qa-FLUO'ROHYDROCORTIS'ONE A solution was prepared containing 320 milligrams of 501,1 15,1701,21-tetrahydroxy-6wmethyl' 9a fluoroallopregnane-3,20-dione, dissolved in eighty milliliters of ethanol denatured with methanol. This solution was purged'with oxygen-free nitrogen gas for a period of ten minutes, and thereto was added a similar oxygen purged solution of 3.3 milliliters of 0.1 normal sodium hydrox EXAMPLE 14 p v 1-dehydr0-6qt-methyl-9a-fluorohydrocortisone (tie-methyl- Three -milliliter portions at a medium, in '25smilliliter Erlenmeyer flasks, containing one percent glucose, two percent corn steep liquor (sixty percent solids) and tap water, were adjusted to a pH of 4.9. This medium was sterilized for 45 minutes at fifteen pounds per square inch pressure and inoculated with a one to two day growth of Sepromyxa aflinis A.T.C.C. 6737. The Erlenmeyer flask was shaken at room temperature (about 26 to 28 degrees centigrade) for a period of three days. At the end of this period this 300-milliliter volume was used as an inoculum for five liters of the same glucose-corn steep liquor medium which in addition contained five milliliters of an antifoam compound (a mixture of lard oil and octadccanol). The fermentor was placed into the water-bath, adjusted to 28 degrees centigrade and the contents stirred (300 r.p.m.) and aerated (0.3 liter air to five liters of beer). After twenty hours of incubation, when agood growth had been developed, one gram of 6a-methyl-9u-fiuorohydrocortisone plus one half gram of 3-ketobisnor-4-cholen-22-al, dissolved in sixteen milliliters of dimethylformamide was added and the incubation carried out at the same temperature (28 degrees centigrade) and aeration for a period of 72 hours (final pH 8.3). The mycelium was filtered off and extracted with three ZOO-milliliter portions of acetone. The beer was extracted with three one-liter portions of methylene chloride and thereupon the extracts of the beer and acetone were combined, dried over anhydrous sodium sulfate and evaporated and the resulting residue chromatographed over a Florisil anhydrous mag nesium silicate column. The column was packed with 100 grams of Florisil and was developed with five 200- milliliter fractions each of methylene chloride, Skelly solve B hexane-acetone mixtures of 9:1, 8:2, 7:3, 1:1, and methanol. The fraction eluted with Skellysolve B acetone (8:2) was twice recrystallized from acetone to give 315 milligrams of 6a-methyl-9a-fiuoro-11fl,17a,21-

trihydroxy-l,4-pregnadiene-3,20-dione of melting point 243 to 250 degrees centigrade with decomposition and rotation plus 93 degrees in acetone.

" A nalysis.--Calcd. for C I-1 1 5 C, 67.33; H, 7.45;

um, Listeria, Erysipelothrix, Mycobacterium, Tritothecium, Leptosphaeria, Cucurbitaria, Nocardia, and enzymes of fungi of the family Tuberculariaceae can be used to introduce a A -bond into -6-methyl 9a fiuorohydrocortisone.

Instead of 6-methyl 9u-fiuorohydrocortisone, ZI-esters thereof can be used such as the .21-acetate, propionate, butyrate, isobutyrate, and the like. Howevenin these cases the ester group is generally saponified during the fermentation process. j

EXAMPLE 15 1 dehydm 6c: methyl 9a-fluorohydrocortisone acetate from fiat-methyl-9a-flu0r0hydrocortisone acetate A mixture of 100 milligrams .of 6m-methyl 9a-fluorohydrocortisone acetate, dissolved in six .milliliters of tertiary butyl alcohol and. 0.55 milliliter of acetic acid was heated together with thirty milligrams of selenium dioxide to 75 degrees centigrade under stirring for a period of 24 hours. Thereafter another thirty-milligram portion of selenium dioxide was added and the mixture heated to 75 degrees centigrade under continuous stirring for a further period of 24 hours. The mixture was then cooled, filtered to remove the selenium dioxide and evaporated. The residue was recrystallized from acetone Skellysolve B four times to give pure 1-dehydro-6a-meth- 20 yl-9'e-f1uorohydrocortisone acetate of melting point237 to 239 degrees centigrade and'rotation [al plus 87 degrees'in acetone. 'iThe infrare'd absorption measured in Nujol mineral oil was' asfollowsf hydroxyl, 3430 cmr' 21-acetoxy-20-keto, 1735, 1717 cm.'- conjugated 3- keto group,.1658-cm.- A -double bonds, 1615, 1610 emsacetate C-O-bond, 1270, 1239 cmr' Analysis.-Calcd. for C H O F: C, 66.34; H, 7.19;

F, 4.37. Found: C, 65.94; H, 6.95; F, 4.72.

7 It is to be understood that the invention is not to be limited to the exact details of operation or exact compounds shown and described, as obvious modifications and equivalents will be apparent to one skilled in the art, and the invention is therefore to be limited only by the scope of the appended claims.

I. claim:

1. A steroid. selected from. the group consisting of 6- methyl-9oc-fluorohydrocortisone, 6-methyl-9a-fluorocortisone, 6-methyl-9u-fluorohydrocortisone 21-acylate and 6- methyl-9m-fluoroc'ortisone 21-acylate wherein the acyl group is the acyl radical of a hydrocarbon carboxylic acid containing from one to twelve carbon atoms, inclusive. 2. 6u-methyl-9a-fluorohydrocortisone.

3. 6m-methyl-9a-fiuorohydrocortisone 21-acetate.

4. 6a-methy1-9u-fluorocortisone.

5. 6a-methyl-9a-fiuorocortisone' 21-acetate.

References Cited in the file of this patent UNITED STATES PATENTS 2,409,798 Reichstein Oct. 22, 1946 2,728,783 Graber Dec. 27, 1955 2,732,383 Bernstein Ian. 24, 1956 2,842,568 Herz .L July 8, 1958 2,852,511 Fried Sept. 16, 1958 OTHER REFERENCES 7 Fried et aL: J.A.C.S. 76,, pages 1455-1456, March 5, 1954. I

Hogg 'et a1.: J.A.C.S. 77, pages 6401-6402, Dec. 5,

UNITED STATES PAiENT OFFICE QERTIFICATE or e Patent No 2 928 85l March 15 1960 George B o Spero Column 2, line 18 for the numeral "37"" read 35 column 5 line 13, for "filtration" read filtration column 12, line 66, for "'conatining read containing column 14 line 52, for "900 milligrams read 200 milligrams column l6 line 9 strike out "and 6-=methyl=-9 -halohydrocortisone"a Signed and sealed this 6th day of September 1960e (SEAL) Attest:

ERNEST W. SWIDER ROBERT C. WATSON Attesting ()fiicer Commissioner of Patents 

1. A STEROID SELECTED FROM THE GROUP CONSISTING OF 6METHYL-9A-FLUOROHYDROCTISONE, 6-METHYL-9A-FLUOROCORTISONE, 6-METHYL-9A-FLUOROHYDROCORTISONE 21-ACYLATE AND 6METHYL-9A-FLUOROCORTISONE 21-ACYLATE WHEREIN THE ACYL GROUP IS THE ACYL RADICAL OF A HYDROCARBON CARBOXYLIC ACID CONTAINING FROM ONE TO TWELVE CARBON ATOMS, INCLUSIVE. 